A comparison of Listeria monocytogenes serovar 4b isolates of clinical and food origin in Japan by pulsed-field gel electrophoresis
Introduction
Outbreaks of foodborne listeriosis were first recognized during the 1980s (Schlech et al., 1983, Fleming et al., 1985, McLauchlin et al., 1991, Jacquet et al., 1995), and it is now known that the consumption of contaminated food is the primary vehicle of transmission (Rocourt and Cossart, 1997, McLauchlin, 1997). Human listeriosis was first recognized in Japan in 1958 (Ito et al., 1959), and although 20–60 sporadic cases are reported every year, specific food vehicles of infection have not been identified.
L. monocytogenes serovar 4b accounts for 60% of the reported human listeriosis in Japan, and strains of serovar 1/2b are responsible for about 30% (Terao, 1991); The predominance of serovar 4b strains is similar to that observed in other countries (McLauchlin, 1990, Farber and Peterkin, 1991). Surveys of food in Japan have shown that among the L. monocytogenes recovered from ready-to-eat foods and raw meat, 25 and 15%, respectively, were serovar 4b (Kokubo et al., 1992; Nakama, unpublished observation). In this study, molecular typing using pulsed-field gel electrophoresis (PFGE) was used to compare L. monocytogenes serovar 4b isolates from listeriosis patients and foods of Japanese origin together with isolates from foodborne incidents which have occurred in North America and Europe.
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Bacterial cultures
A total of 120 Listeria monocytogenes serovar 4b isolates were selected for study and were comprised of 82 isolates from listeriosis patients in Japan between 1978 and 1993 (including three cases of congenital infection); 20 isolates from dairy, meat or seafood products on retail sale in Japan between 1989 and 1993; 16 isolates from eight incidents of foodborne listeriosis in North America and Europe (Schlech et al., 1983, Bannister, 1987, Linnan et al., 1988, Azadian et al., 1989, Bille, 1990,
Results
PFGE of genomic DNA after digestion with SmaI or ApaI yielded 10–15 or 11–16 fragments, respectively, which ranged in size from 50 to 500 kb (Fig. 1a,b). Digestion with AscI or Sse8387I produced banding patterns both comprising six to nine fragments ranging in size from 50 kb to 1 Mb (Fig. 1c,d). The PFGE profiles derived from all 120 cultures produced 18 patterns with SmaI, 22 with ApaI, 22 with AscI, and 15 with Sse8387I. A combination of PFGE profiles of all four enzymes produced 60 distinct
Discussion
Eighty-two isolates of L. monocytogenes serovar 4b recovered from listeriosis patients in Japan and tested by PFGE using four restriction enzymes (SmaI, ApaI, AscI and Sse8387I) showed a predominant PFGE type which was indistinguishable from that responsible for two large foodborne outbreaks in California (1985) and Switzerland (1983–1987).
Buchrieser et al. (1993)analyzed cultures from foodborne listeriosis by PFGE using ApaI, SmaI and NotI, and reported that the isolates from the major
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